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亚洲梨病原菌梨火疫病菌荚膜胞外多糖生物合成与结构的遗传学研究

Genetics of biosynthesis and structure of the capsular exopolysaccharide from the Asian pear pathogen Erwinia pyrifoliae.

作者信息

Kim Won-Sik, Schollmeyer Martin, Nimtz Manfred, Wray Victor, Geider Klaus

机构信息

Max-Planck-Institut für Zellbiologie, Rosenhof, 68526 Ladenburg, Germany1.

GBF, Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, 38124 Braunschweig, Germany2.

出版信息

Microbiology (Reading). 2002 Dec;148(Pt 12):4015-4024. doi: 10.1099/00221287-148-12-4015.

Abstract

Erwinia pyrifoliae is a novel bacterial pathogen, which causes Asian pear blight and is related to Erwinia amylovora, the causative agent of fire blight. E. pyrifoliae produces exopolysaccharide (EPS) related to amylovoran in its sugar composition and sugar linkages. This was shown by degradation of the EPS with a viral depolymerase, and by methylation analysis and ESI/MS. The structure of the repeating units was confirmed by (1)H-NMR spectra. The EPS of E. pyrifoliae carried side chains, which were mainly terminated by acetyl and pyruvyl residues as found previously for amylovoran. On the other hand, a second side chain with glucose found for up to 65% of the repeating units of amylovoran was completely absent. The nucleotide sequences of five genes of the cps cluster of E. pyrifoliae encoding proteins for EPS synthesis were characterized and displayed a high homology with the corresponding ams genes. Similar functions of the gene products are assumed. As for ams mutants of E. amylovora, a cpsB mutant of E. pyrifoliae did not synthesize EPS and did not produce ooze on slices of immature pears or symptoms on pear seedlings. The cps mutant was complemented for EPS synthesis and virulence on pear slices with a gene cluster of E. amylovora that included amsB.

摘要

梨火疫欧文氏菌是一种新型细菌病原体,可引起亚洲梨疫病,与梨火疫病的病原体梨火疫欧文氏菌有关。梨火疫欧文氏菌产生的胞外多糖(EPS)在糖组成和糖键方面与淀粉葡聚糖有关。这通过用病毒解聚酶降解EPS、甲基化分析和电喷雾电离质谱(ESI/MS)得以证明。重复单元的结构通过氢核磁共振谱(¹H-NMR)得以确认。梨火疫欧文氏菌的EPS带有侧链,这些侧链主要由乙酰基和丙酮酰基残基终止,这与之前在淀粉葡聚糖中发现的情况相同。另一方面,在高达65%的淀粉葡聚糖重复单元中发现的带有葡萄糖的第二条侧链则完全不存在。对梨火疫欧文氏菌编码EPS合成蛋白的cps基因簇的五个基因的核苷酸序列进行了表征,并显示与相应的ams基因具有高度同源性。推测基因产物具有相似功能。对于梨火疫欧文氏菌的ams突变体,梨火疫欧文氏菌的cpsB突变体不合成EPS,在未成熟梨切片上不产生菌脓,在梨幼苗上也不产生症状。用包含amsB的梨火疫欧文氏菌基因簇对cps突变体进行补充,使其在梨切片上恢复EPS合成和毒力。

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